Integrative Metabolomics and Proteomics, Berlin Institute of Medical Systems Biology/Max-Delbrueck Center for Molecular Medicine, Robert Rossle Street 10, Berlin 13125, Germany.
Present address: Faculté des Sciences, de la Technologie et de la Communication, University of Luxembourg, 162 A, Avenue de la Faïencerie L-1511, Luxembourg, Luxembourg.
Cancer Metab. 2014 Jun 30;2:9. doi: 10.1186/2049-3002-2-9. eCollection 2014.
Cellular metabolism is highly dynamic and continuously adjusts to the physiological program of the cell. The regulation of metabolism appears at all biological levels: (post-) transcriptional, (post-) translational, and allosteric. This regulatory information is expressed in the metabolome, but in a complex manner. To decode such complex information, new methods are needed in order to facilitate dynamic metabolic characterization at high resolution.
Here, we describe pulsed stable isotope-resolved metabolomics (pSIRM) as a tool for the dynamic metabolic characterization of cellular metabolism. We have adapted gas chromatography-coupled mass spectrometric methods for metabolomic profiling and stable isotope-resolved metabolomics. In addition, we have improved robustness and reproducibility and implemented a strategy for the absolute quantification of metabolites.
By way of examples, we have applied this methodology to characterize central carbon metabolism of a panel of cancer cell lines and to determine the mode of metabolic inhibition of glycolytic inhibitors in times ranging from minutes to hours. Using pSIRM, we observed that 2-deoxyglucose is a metabolic inhibitor, but does not directly act on the glycolytic cascade.
细胞代谢具有高度动态性，并不断适应细胞的生理程序。代谢的调节出现在所有的生物层面：（后）转录、（后）翻译和变构。这种调节信息在代谢组中表现出来，但方式复杂。为了解码这种复杂的信息，需要新的方法来促进高分辨率的动态代谢特征描述。
在这里，我们将脉冲稳定同位素分辨代谢组学（pSIRM）描述为一种用于细胞代谢动态代谢特征描述的工具。我们已经适应了气相色谱-质谱联用方法进行代谢组学分析和稳定同位素分辨代谢组学。此外，我们还提高了稳健性和重现性，并实施了一种用于代谢物绝对定量的策略。
通过实例，我们应用该方法来描述一系列癌细胞系的中心碳代谢，并确定糖酵解抑制剂在从分钟到小时的时间范围内的代谢抑制模式。使用 pSIRM，我们观察到 2-脱氧葡萄糖是一种代谢抑制剂，但不会直接作用于糖酵解级联。
